I need to do hybridisations with probes of about 100 bp and 20 bp on
heterologous target dna's. The expected HOMOLOGY between the probes and the
target is about 60%. I realise that the 20 bp probe probably won't work but
there has to be a nice PROTOCOL for the 100 bp probe. I have to do
this by colony hybridisation, so I will have a lot DNA on my blots (HYBOND N
nylon membranes (AMERSHAM)). As a possitive control I will use the homologous
target. Is it possible to discriminate the homologous target and the
heterologous target just by using two consecutive washing steps?
DOES ANYONE HAS A NICE HETEROLOGOUS HYBRIDISATION PROTOCOL FOR ME ?
OR IS THERE SOMOWHERE ON INTERNET A LIBRARY WHICH CONTAINS THIS KIND OF
PROTOCOLS?
THANK YOU IN ADVANCE FOR YOUR REPLY
MARC VAN DE CRAEN
E-MAIL : MARCVDC at LMB1.RUG.AC.BE